Primary cells undergo a finite number of population doublings and reach a state called replicative senescence after limited cell divisions. GeneCopoeia’s Cell Immortalization Reagents use recombinant lentivirus carrying genes, such as the simian virus 40 (SV40) T antigen, the human telomerase reverse transcriptase protein (hTERT), CDK4, HOXB, HOXA, cMyc, Bmi1 and HPV-16 E6/E7 to induce immortalization and significantly prolong primary cells’ lifespan.
Product details
GeneCopoeia Lentifect™ Lentiviral Particles are produced from a standardized protocol using purified plasmid DNA and the proprietary reagents EndoFectin™ Lenti (for transfection) and TiterBoost™ solution. The protocol uses a third-generation self-inactivating packaging system meeting BioSafety Level 2 requirements.
The Lentifect™ particles include CMV or EF1α promoter for efficient expression of the immortalization inducer gene in target cells with puromycin or hygromycin resistance marker for selection of stably transduced cells. The immortalization lentivirus particles without antibiotic selection marker are also available.
Provided as 4 vials of 25 µl of lentiviral particles with titers of ~1 x 108 TU/ml. Lentifect™ particles are shipped on dry ice and must be stored at –80°C immediately upon receipt. Avoid repeated freeze-thaw cycles as this will reduce titers.
The lentiviral expression construct was validated by full-length sequencing, restriction enzyme digestion and PCR-size validation using gene-specific and vector-specific primers. The product is confirmed free of bacteria, fungi and common Mycoplasma contamination.
